This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Use FCS to determine the concentrations, aggregation states, and interactions of the following molecules (in priority order) in migrating cells. (paxillin, a-actinin;FAK and paxillin;GIT1 and paxillin;GIT1 and Pix, Rac, or Pak). Determine concentrations and degree of aggregation by FCS. (1) number of molecules in and around adhesions;(2) ectopic expression and then endogenous expression levels;(3) detrmine the number of complexed molecules in and around adhesions;(4) detect activation of soluble components by FCS (paK, Rac);(5) determine changes in aggregation and interactions as adhesions form and disperse. and determine dynamics of single and interacting proteins using FCS